Laboratory of Clinical Pharmacology, Jiaozhou Central Hospital, Qingdao, Shandong Province, China.
Department of Information, Jiaozhou Central Hospital, Qingdao, Shandong Province, China.
Biomed Chromatogr. 2021 Apr;35(4):e5031. doi: 10.1002/bmc.5031. Epub 2020 Dec 3.
This study aimed to develop a highly selective, sensitive and fast liquid chromatography tandem mass spectrometric (LC-MS/MS) method for the determination of obacunone in rat plasma. Sample preparation was accomplished by a simple solid-phase extraction procedure. Chromatographic separation was carried out on an ACQUITY BEH C column using acetonitrile/methanol (1:1, v/v) and 0.1% formic acid in water as mobile phase at a flow rate of 0.4 mL/min. Quantification was performed with multiple reactions monitoring in positive ion mode with the precursor-to-product ion transitions at m/z 455.2 > 161.1 for obacunone and m/z 515.2 > 161.1 for nomilin (internal standard). The assay was demonstrated to be linear over the concentration range of 0.1-1,000 ng/mL with correlation coefficient >0.999 (r > 0.999). The intra- and inter-day accuracy ranged from -8.33 to 10.40%, while the precision was <10.41%. The mean extraction recovery was >75.32%, and the assay was free of matrix effect. The validated LC-MS/MS method was successfully applied to the pharmacokinetic study of obacunone in rats after oral and intravenous administrations. The oral bioavailability of obacunone was 13.59%.
本研究旨在开发一种高选择性、高灵敏度、快速的液相色谱串联质谱（LC-MS/MS）方法，用于测定大鼠血浆中的橙酮。样品制备采用简单的固相萃取程序完成。采用 ACQUITY BEH C 柱，以乙腈/甲醇（1:1，v/v）和 0.1%甲酸水溶液为流动相，流速为 0.4 mL/min，进行色谱分离。采用正离子模式下的多重反应监测进行定量分析，橙酮的前体-产物离子跃迁为 m/z 455.2 > 161.1，诺米林（内标）的前体-产物离子跃迁为 m/z 515.2 > 161.1。该测定方法在 0.1-1000ng/mL 的浓度范围内呈线性，相关系数>0.999（r>0.999）。日内和日间准确度的范围为-8.33%至 10.40%，而精密度<10.41%。平均提取回收率>75.32%，且该测定方法无基质效应。经验证的 LC-MS/MS 方法成功应用于大鼠口服和静脉给药后橙酮的药代动力学研究。橙酮的口服生物利用度为 13.59%。
